GSM7049306: E77 Occipital DIV8; Macaca mulatta; RNA-Seq
Total RNA was extracted from 2 different wells/condition using RNeasy Mini Kit (QIAGEN, 74104), according to manufacturer's protocol. Total RNA quality was determined by estimating the A260/A280 and A260/A230 ratios by nanodrop. RNA integrity was determined by running an Agilent Bioanalyzer gel. mRNA was purified from approximately 200ng of total RNA with oligo-dT beads and sheared by incubation at 94C in  presence of Mg (Roche Kapa mRNA Hyper Prep Cat# KR1352).  Following first-strand synthesis with random primers, second strand synthesis and A-tailing were performed with dUTP for generating strand-specific sequencing libraries. Indexed libraries were quantified by qRT-PCR using a kit (Roch KAPA Biosystems Cat# KK4854) and insert size distribution determined by either the Agilent Bioanalyzer. Samples with a yield of ≥ 0.5 ng/ul and a size distribution of 150-300bp were used for sequencing.